镁合金通过调控PI3K/AKT信号通路促进巨噬细胞M2极化在肩袖损伤修复中肌腱-骨愈合作用的分子机制研究  

Molecular mechanism of magnesium alloy promoting macrophage M2 polarization through modulation of PI3K/AKT signaling pathway for tendon-bone healing in rotator cuff injury repair

作　　者：盛显浩 张文 宋首龙 张飞 张宝祥 田晓莹 熊文韬 朱颖广 谢雨欣 李子昂 谭丽丽[3] 张强 王岩 SHENG Xianhao;ZHANG Wen;SONG Shoulong;ZHANG Fei;ZHANG Baoxiang;TIAN Xiaoying;XIONG Wentao;ZHU Yingguang;XIE Yuxin;LI Zi’ang;TAN Lili;ZHANG Qiang;WANG Yan(Chinese PLA Medical School,Beijing,100853,P.R.China;Department of Orthopedics,the Fourth Medical Center of Chinese PLA General Hospital,Beijing,100048,P.R.China;Institute of Metal Research,Chinese Academy of Sciences,Shenyang Liaoning,110013,P.R.China)

机构地区：[1]中国人民解放军医学院,北京100853 [2]中国人民解放军总医院第四医学中心骨科医学部,北京100048 [3]中国科学院金属研究所,沈阳110013

出　　处：《中国修复重建外科杂志》2025年第2期174-186,共13页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：首都卫生发展科研专项项目(首发2022-2-5051)。

摘　　要：目的评估生物可降解镁合金材料在肩袖撕裂修复中促进肌腱-骨愈合的效果,并探讨其潜在的生物学机制。方法取8周龄SD大鼠48只,随机分为A、B、C 3组。A、B组大鼠制备肩袖撕裂模型后分别通过镁合金缝线或Vicryl Plus 4-0可吸收缝线进行修复,C组仅行皮下切开和缝合。术后1、2周取A、B组内脏行HE染色评估镁合金的安全性;术后2、4、8、12周取3组冈上肌腱-肱骨近端标本,其中4、12周标本行大体观察,4、8、12周标本行生物力学检测A、B组愈合部位的极限载荷及刚度;2、4、12周标本行以下检测:Micro-CT评估A、B组骨隧道形成情况,标本脱钙切片后行HE染色和Masson染色观察肌腱-骨界面纤维软骨再生情况,Goldner三色染色评估钙化情况,免疫组织化学染色检测肌腱-骨界面血管VEGF、BMP-2等成血管因子及成骨因子的表达,免疫荧光染色通过检测精氨酸酶1、整合素β2亚基观察肌腱-骨界面巨噬细胞M1、M2极化情况,通过实时荧光定量PCR分析磷脂酰肌醇3激酶(phosphatidylinositede 3-kinase,PI3K)/蛋白激酶B(protein kinase B,AKT)信号通路在肌腱-骨愈合中的作用机制。结果内脏组织HE染色示,镁合金降解过程中释放的镁离子对心脏、肝脏、脾脏、肺、肾脏等器官未产生明显毒性作用,其生物安全性良好。冈上肌腱-肱骨近端标本组织学染色示,A组肌腱-骨界面处的纤维软骨再生恢复较早,且纤维软骨数量明显多于B组,提示镁合金材料对肌腱-骨界面修复具有积极作用。Micro-CT分析示,A组骨隧道形成快于B组,进一步验证了镁合金材料对骨愈合的促进作用。生物力学测试示,A组的极限载荷均高于B组,4周时A组刚度高于B组,表明A组肌腱-骨界面修复后的组织承载能力更强,镁合金具有改善肌腱-骨愈合的潜力。免疫组织化学染色示,肌腱-骨愈合早期VEGF和BMP-2表达显著上调,提示镁合金能够有效促进血管生成与骨形成Objective To evaluate the effect of biodegradable magnesium alloy materials in promoting tendonbone healing during rotator cuff tear repair and to investigate their potential underlying biological mechanisms.Methods Forty-eight 8-week-old Sprague Dawley rats were taken and randomly divided into groups A,B,and C.Rotator cuff tear models were created and repaired using magnesium alloy sutures in group A and Vicryl Plus 4-0 absorbable sutures in group B,while only subcutaneous incisions and sutures were performed in group C.Organ samples of groups A and B were taken for HE staining at 1 and 2 weeks after operation to evaluate the safety of magnesium alloy,and specimens from the supraspinatus tendon and proximal humerus were harvested at 2,4,8,and 12 weeks after operation.The specimens were observed macroscopically at 4 and 12 weeks after operation.Biomechanical tests were performed at 4,8,and 12 weeks to test the ultimate load and stiffness of the healing sites in groups A and B.At 2,4,and 12 weeks,the specimens were subjected to the following tests:Micro-CT to evaluate the formation of bone tunnels in groups A and B,HE staining and Masson staining to observe the regeneration of fibrocartilage at the tendon-bone interface after decalcification and sectioning,and Goldner trichrome staining to evaluate the calcification.Immunohistochemical staining was performed to detect the expressions of angiogenic factors,including vascular endothelial growth factor(VEGF)and bone morphogenetic protein 2(BMP-2),as well as osteogenic factors at the tendon-bone interface.Additionally,immunofluorescence staining was used to examine the expressions of Arginase 1 and Integrin beta-2 to assess M1 and M2 macrophage polarization at the tendon-bone interface.The role of the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway in tendon-bone healing was further analyzed using real-time fluorescence quantitative PCR.Results Analysis of visceral sections revealed that magnesium ions released during the degradation of magn

关 键 词：镁合金 肩袖撕裂 肌腱-骨愈合 磷脂酰肌醇3激酶 蛋白激酶B 信号通路 巨噬细胞 

分 类 号：R686[医药卫生—骨科学]